Recombinant Mouse TNFRSF19/TROY

Using Recombinant Mouse TNFRSF19/TROY in research applications is valuable for studying its roles in cell signaling, cancer biology, stem cell function, and developmental processes. This recombinant protein enables controlled, reproducible experiments to dissect TROY’s molecular mechanisms and biological effects.

Key scientific reasons to use recombinant mouse TNFRSF19/TROY include:

• Cancer Research: TROY is overexpressed in glioblastoma and promotes tumor cell migration, invasion, and resistance to therapies by activating survival pathways such as Akt and NF-κB. Recombinant TROY allows for in vitro and in vivo studies to:

  • Investigate how TROY signaling contributes to tumor progression and therapeutic resistance.
  • Screen for inhibitors or antibodies that block TROY-mediated effects.
  • Model TROY-driven phenotypes in cell lines or animal models.

• Stem Cell and Developmental Biology: TROY marks basal cells in the interfollicular epidermis and hair follicle infundibulum, identifying populations with stem cell properties. Recombinant TROY can be used to:

  • Study its role in epidermal stem cell maintenance, differentiation, and tissue regeneration.
  • Perform organoid assays to assess stem cell capacity and lineage tracing.

• Cell Death and Survival Signaling: TROY can induce apoptosis via caspase-independent mechanisms and modulate cell survival through pathways such as c-Jun and NF-κB. Applications include:

  • Elucidating the balance between cell death and survival in various tissues.
  • Dissecting downstream signaling cascades using biochemical and cell-based assays.

• Developmental Studies: TROY is expressed in multiple murine tissues, especially during late embryonic development, and is implicated in organogenesis. Recombinant protein can be used to:

  • Analyze developmental signaling pathways in embryonic models.
  • Study tissue-specific functions by adding recombinant TROY to primary cell cultures.

• Assay Development and Screening: Recombinant TROY is essential for:

  • Generating standard curves in ELISA or Western blot assays.
  • Validating antibody specificity and sensitivity.
  • High-throughput screening of small molecules or biologics targeting TROY.

• Mechanistic Studies: Using recombinant protein enables:

  • Controlled stimulation of cells to study receptor-ligand interactions.
  • Dissection of TROY’s role in signaling networks, including interactions with JAK1, Pyk2-Rac1, and other pathways.

In summary, recombinant mouse TNFRSF19/TROY is a critical reagent for mechanistic, translational, and therapeutic research in oncology, stem cell biology, and developmental biology, providing a defined and reproducible tool to interrogate TROY’s diverse biological functions.

Yes, recombinant Mouse TNFRSF19/TROY protein can be used as a standard for quantification or calibration in ELISA assays, provided it is validated for this purpose and matches the assay’s requirements. Recombinant standards are commonly used in quantitative ELISA protocols to generate standard curves, enabling accurate measurement of target analyte concentrations in biological samples.

Essential context and supporting details:

• Assay Principle: Quantitative ELISA kits for Mouse TNFRSF19/TROY typically employ a sandwich format, where a monoclonal antibody specific for TNFRSF19/TROY is pre-coated onto the plate. Recombinant Mouse TNFRSF19/TROY is used as the standard to generate a calibration curve, against which sample concentrations are determined.
• Validation: The recombinant standard should be highly purified and its concentration accurately determined. The immunoassay must be calibrated against this recombinant protein, and the standard curve should be generated for each assay run to ensure precision and accuracy.
• Specificity: These ELISA kits are designed to recognize both recombinant and natural Mouse TNFRSF19/TROY, with no significant cross-reactivity reported with other proteins.
• Standard Preparation: The recombinant standard is typically supplied lyophilized and must be reconstituted and serially diluted according to the kit protocol to cover the assay’s dynamic range (e.g., 62.5–4000 pg/mL).
• Carrier Proteins: Some protocols recommend using recombinant protein with a carrier (e.g., BSA) for stability, especially for ELISA applications. Carrier-free preparations may be preferred for other uses, but for ELISA calibration, stability and solubility are critical.

Best practices:

• Always prepare a fresh standard curve for each assay.
• Confirm that the recombinant standard’s sequence and post-translational modifications (if any) are compatible with the antibodies used in your ELISA.
• Validate the standard in your specific sample matrix if you are using it outside the recommended matrices (e.g., serum, plasma, cell culture supernatant).
• Use appropriate diluents and mixing protocols to ensure reproducibility.

Limitations:

• The recombinant standard must be lot-specific and its concentration verified, as variations can affect quantification accuracy.
• If using a recombinant standard not supplied with your ELISA kit, ensure it is compatible with the kit’s antibodies and detection system.

Summary: Recombinant Mouse TNFRSF19/TROY is suitable as a standard for ELISA quantification, provided it is validated and matches the assay’s requirements for specificity, purity, and concentration.

Recombinant Mouse TNFRSF19/TROY has been validated for several key applications in published research, primarily in studies of cell signaling, stem cell biology, and cancer pathobiology.

Validated Applications in Published Research:

• Cellular and Molecular Functional Studies:
  Recombinant TROY has been used to investigate its role in glioblastoma cell survival, migration, and therapeutic resistance. Overexpression and knockdown experiments in mouse models and cell lines have demonstrated that TROY promotes glioblastoma invasion and resistance to apoptosis, particularly through activation of the Akt and NF-κB signaling pathways. These studies often employ recombinant TROY for transfection, overexpression, or as a ligand in signaling assays.

• Stem Cell and Lineage Tracing Studies:
  TROY (Tnfrsf19) has been identified as a marker for specific epidermal and intestinal stem cell populations in mice. Recombinant TROY is used in lineage tracing, organoid culture, and single-cell transcriptomics to study stem cell capacity and tissue regeneration, particularly in the skin and intestinal epithelium.

• In Vitro Assays:
  Recombinant mouse TROY is used as a standard or ligand in ELISA and cell-based assays to study receptor-ligand interactions, downstream signaling, and protein-protein interactions. These applications are supported by the protein’s use in sandwich immunoassays and as a control in biochemical studies.

• In Vivo Functional Studies:
  Recombinant TROY has been utilized in in vivo gene transfer and xenograft models to assess its effects on cell migration, tumor progression, and survival in mouse models of glioblastoma.

Additional Context:

• Detection and Quantification:
  Antibodies against mouse TROY, often validated using recombinant protein, are widely used in Western blotting, ELISA, flow cytometry, and immunocytochemistry to detect TROY expression in tissues and cells.

• Tissue Distribution and Developmental Studies:
  Recombinant TROY has been used to study its expression pattern in various murine tissues, especially in the brain, reproductive organs, and during embryonic development.

Summary Table: Applications of Recombinant Mouse TNFRSF19/TROY

These applications are well-supported in the literature, particularly in the context of cancer biology, stem cell research, and receptor signaling pathways.

To reconstitute and prepare Recombinant Mouse TNFRSF19/TROY protein for cell culture experiments, follow these best-practice steps:

1. Centrifuge the vial briefly to collect all lyophilized protein at the bottom before opening.

2. Reconstitution:

   • Use sterile, 0.2 μm-filtered phosphate-buffered saline (PBS), pH 7.2–7.4, as the reconstitution buffer.
   • For enhanced stability, especially at low concentrations, add at least 0.1% carrier protein (such as bovine serum albumin, BSA, or human serum albumin) to the buffer.
   • Typical reconstitution concentrations range from 0.1 to 1.0 mg/mL. For example, to achieve 0.2 mg/mL, add 500 μL PBS to 100 μg lyophilized protein.
   • Gently pipette up and down or swirl to dissolve. Avoid vigorous vortexing to prevent protein denaturation.

3. Aliquoting and Storage:

   • Once fully dissolved, aliquot the solution into small volumes to avoid repeated freeze-thaw cycles.
   • Store aliquots at –20°C to –80°C for long-term storage. For short-term use (up to 1 month), 2–8°C is acceptable.
   • Avoid multiple freeze-thaw cycles, as this can degrade the protein.

4. Preparation for Cell Culture:

   • Before adding to cell culture, dilute the reconstituted protein to the desired working concentration using cell culture medium or PBS with 0.1% carrier protein.
   • If the protein was reconstituted in PBS with carrier, ensure compatibility with your cell culture system.
   • Filter the final working solution through a 0.2 μm sterile filter if sterility is required.

5. General Notes:

   • Always consult the specific product datasheet for any lot-specific instructions, as formulation and recommended buffers may vary.
   • If the protein contains tags (e.g., Fc, His), confirm that these do not interfere with your experimental system.

Summary Table:

These steps will help ensure protein stability, sterility, and biological activity for cell culture applications.